New Compounds

ABSTRACT

The invention relates to a compound of formula (I) wherein the variables have the meaning as indicated in the claims; in free form and in salt form; and optionally the enantiomers and geometrical isomers thereof. The compounds of formula (I) are useful as therapeutic agent for organ transplants, lupus, multiple sclerosis, rheumatoid arthritis, psoriasis. Type I diabetes and complications from diabetes, cancer, asthma, atopic dermatitis, autoimmune thyroid disorders, ulcerative colitis, inflammatory bowel diseases, Crohn&#39;s disease, Alzheimer&#39;s disease, leukemia, osteoarthritis, control of pruritus, chronic respiratory disease or keratoconjunctivitis in mammals.

The present invention relates to novel heterocyclyl-substitutedcyclohexylmethansulfonamides which are Janus kinase inhibitors, alsoknown as JAK inhibitors, and their use in the treatment of allergicreactions including allergic dermatitis, eczema, atopic dermatitis,pruritus and other pruritic conditions and also inflammatory diseases.

Oclacitinib, a pyrrolopyrimidinaminocyctohexylmethansulfonamide, is aJAK inhibitor, which is approved for the control of pruritus associatedwith allergic dermatitis and the control of atopic dermatitis in dogs.However, the search for new, more potent JAK inhibitor moleculescontinues. Surprisingly, new specific JAK inhibitors have been foundwhich provide an improved activity concerning skin diseases, inparticular atopic dermatitis and pruritus.

The present invention therefore in one aspect concerns a compound offormula

wherein R is C₁-C₄-alkyl or C₃-C₅-cycloalkyl; and A is

(i) a radical of formula

wherein either Y is NH, N(C₁-C₂-alkyl) or O, R² is cyano (CN), nitro(NO₂) C(O)NR′R″, C(O)OR′ or NR′R″, and R′ and R″ are each independentlyof the other H or C₁-C₄-alkyl; or

Y and R² together form a bivalent radical

wherein R⁰ is H, C₁-C₄-alkyl, hydroxy-C₁-C₄-alkyl,C₁-C₄-alkoxy-C₁-C₄-alkyl or NH₂ and preferably H, C₁-C₄-alkyl or NH₂;or is (ii) a radical of formula

wherein X is N, C(CN), C(NO₂), C[C(O)NR′R″], C[C(O)OR′] or C(NR′R″), andY, R′ and R″ are as defined above;

or is (iii) a radical of formula

wherein one of X¹ and X² is N and the other one is C(R³), R³ is H,C₁-C₄-alkyl, phenyl or benzyl, and Y is as defined above;

or is (iv) a radical of formula

wherein either X³ is N and X⁴ is CH or N, or X¹ is C(CN), C(NO₂),C[C(O)NR′R″], C[C(O)OR′] or C(NR′R″) and X⁴ is N, and wherein Y. R′ andR″ are as defined above;

or is (v) a radical of formula

wherein R⁴ and R⁵ are each independently of the other H, halogen,C₁-C₄-alkyl or phenyl, and Y is as defined above.

The variable R is preferably methyl, ethyl or cyclobutyl, in particularmethyl.

R⁰ as hydroxyl-C₁-C₄-alkyl is preferably hydroxymethyl or hydroxyethyl,in particular hydroxymethyl. R⁰ as C₁-C₄-alkoxy-C₁-C₄-alkyl ispreferably methoxymethyl or ethoxymethyl, in particular methoxymethyl.The variable R⁰ is preferably H, methyl hydroxymethyl, methoxymethyl orNH₂, more preferably H, methyl or NH₂, in particular H. R′ and R″ areeach independently of the other preferred H, methyl or ethyl. R² ispreferably cyano or nitro.

One embodiment of the invention concerns a radical A of formula (2a),wherein R² and Y have the meanings as defined. A preferred radical A isof formula (2a), wherein R² is cyano or nitro and Y is NH, N(CH₃) or O,preferably NH or N(CH₃), in particular N(CH₃).

Still a further preferred radical A is a radical of formula

Preferred radicals of formula (2b) are a radical

wherein Y is in each case O, NH or N(CH₃), in particular N(CH₃), or O,in particular N(CH₃).

A further embodiment of the invention concerns a radical A of formula(2c), wherein X¹ is CH and X² is N. Still a further embodiment concernsa radical A of formula (2c) wherein X¹ is N, X² is C(R³) and R³ is H,C₁-C₄-alkyl, phenyl or benzyl, preferably H, methyl, phenyl or benzyl.

Preferred radicals of formula (2d) are:

wherein Y is in each case NH or N(CH₃).

A further embodiment concerns a radical A of formula (2e), wherein oneof R⁴ and R⁵ is H and the other one is H, halogen, or phenyl and Y is NHor N(CH₃).

The compounds of this invention can exist as one or more stereoisomers.The various stereoisomers include enantiomers, diastereomers,atropisomers and geometric isomers.

In case the compounds of the formula (I) have a chiral carbon atom, theymay have either an (R) or an (S) configuration. The present inventionencompasses compounds formula (I) both with (S) and with (R)configuration at the particular chiral carbon atoms, which means thatthe present invention covers the compounds of the general formula (I) inwhich the carbon atoms in question each independently have an (R)configuration; or have an (S) configuration.

If a plurality of chiral centres are present in the compounds of theformula (I) any desired combinations of the configurations of the chiralcentres are possible, which means that (1) one chiral centre may have(R) configuration and the other chiral centre (S) configuration; (2) onechiral centre may have (R) configuration and the other chiral centre (R)configuration; and (3) one chiral centre may have (S) configuration andthe other chiral centre (S) configuration.

One skilled in the art will appreciate that one stereoisomer may be moreactive and/or may exhibit beneficial effects when enriched relative tothe other stereoisomers) or when separated from the otherstereoisomers). Additionally, the skilled artisan knows how to separate,enrich, and/or to selectively prepare said stereoisomers. The compoundsof the invention may be present as a mixture of stereoisomers,individual stereoisomers, or as an optically active form.

One skilled in the art will appreciate that not all nitrogen containingheterocyclic rings can form N-oxides since the nitrogen requires anavailable lone pair for oxidation to the oxide; one skilled in the artwill recognize those nitrogen containing heterocyclic rings which canform N-oxides. One skilled in the art will also recognize that tertiaryamines can form N-oxides. Synthetic methods for the preparation ofN-oxides of heterocyclic rings and tertiary amines are very well knownby one skilled in the art including the oxidation of heterocyclic ringsand tertiary amines with peroxy acids such as peracetic andm-chloroperbenzoic acid (MCPBA), hydrogen peroxide, alkyl hydroperoxidessuch as t-butyl hydroperoxide, sodium perborate, and dioxiranes such asdimethyl dioxirane These methods for the preparation of N-oxides havebeen extensively described and reviewed in the literature. Themanufacture of suitable S-oxides may be performed in an analogous mannerusing, for example, the same kind of oxidants as mentioned above for theN-oxides.

One skilled m the art recognizes that because of the environment andunder physiological conditions salts of chemical compounds are inequilibrium with their corresponding nonsalt forms, salts share thebiological utility of the nonsalt forms. Thus a wide variety of salts ofthe compounds of formula (I) are useful (i.e. are veterinarilysuitable). The salts of the compounds of formula (I) includeacid-addition salts with inorganic or organic acids such as hydrobromic,hydrochloric, nitric, phosphoric, sulfuric, acetic, butyric, fumaric,lactic, maleic, malonic, oxalic, propionic, salicylic, tartaric,4-toluenesulfonic “or valeric acids. When a compound of formula (I)contains an acidic moiety such as a carboxylic acid or phenol, saltsalso include those formed with organic or inorganic bases such aspyridine, triethylamine or ammonia, or amides, hydrides, hydroxides orcarbonates of sodium, potassium, lithium, calcium, magnesium or barium.Accordingly, the present invention comprises compounds selected fromformula (I), N-oxides and veterinary acceptable salts thereof. Thecompounds of the present invention can also form internal salts.

The present invention also provides pro-drugs of the compounds of thepresent invention that converts in vivo to the compounds of the presentinvention. A pro-drug is an active or inactive compound that is modifiedchemically through in vivo physiological action, such as hydrolysis,metabolism and the like, into a compound of this invention followingadministration of the prodrug to a subject. Exemplary prodrugs are,e.g., esters of free carboxylic acids and S-acyl and O-acyl derivativesof thiols, alcohols or phenols, wherein acyl has a meaning as definedherein. Preferred are veterinary or pharmaceutically acceptable esterderivatives convertible by solvolysis under physiological conditions tothe parent carboxylic acid, e.g., lower alkyl esters, cycloalkyl esters,lower alkenyl esters, benzyl esters, mono- or di-substituted lower alkylesters, such as the -(amino, mono- or di-lower alkylamino, carboxy,lower alkoxycarbonyl)-lower alkyl esters, the -(lower alkanoyloxy, loweralkoxycarbonyl or di-lower alkylaminocarbonyl)-lower alkyl esters, suchas the pivaloyloxymethyl ester and the like conventionally used in theart. In addition, amines have been masked as arylcarbonyloxymethylsubstituted derivatives which are cleaved by esterases in vivo releasingthe free drug and formaldehyde. Moreover, drugs containing an acidic NHgroup, such as imidazole, imide, indole and the like, have been maskedwith N-acyloxy-methyl groups. Hydroxy groups have been masked as estersand ethers. EP0 039 051 discloses Mannich-base hydroxamic acid prodrugs,their preparation and use. In view of the close relationship between thecompounds, the compounds in the form of their salts and the pro-drugs,any reference to the compounds of the present invention is to beunderstood as referring also to the corresponding pro-drugs of thecompounds of the present invention, as appropriate and expedient.

The compounds of formula (I) may be prepared, for example, by reacting acompound of

Formula

A′-Hal  (3) with

a compound of formula

wherein Y and R are as defined above, Hal is halogen, for example,chlorine, and A′ is a radical of formula

wherein R², R⁴, R⁵, X¹, X², X³ and X⁴ are as defined above. Thenucleophilic substitution reaction may be performed, for example, asdescribed in textbooks of organic chemistry. For example, the compoundsof formula (3) and (4) are reacted in a suitable solvent or mixture ofsolvents in the presence of a base. The choice of solvent and base isstrongly dependent on the specific nature of the compounds of formulae(3) and (4). The reaction may take place at room temperature or atelevated temperature, for example, above 100° C. In case of a compoundof formula (3) comprising a radical A′ with a NH group, it may beadvisable to protect said amine group before performing the reactionwith the compound of formula (4). The protection of the amine group anddeprotection afterwards may be performed in a manner known per se.

Alternatively the compounds of formula (3) and (4) can be coupled viathe Buchwald-Hartwig Pd catalyzed amination as described in textbooks oforganic chemistry.

The compounds of formula (3) are known per se or may be preparedaccording to methods known per sc.

The compounds of formula (4) are likewise known, or may be preparedaccording to methods known per sc. For example, the HY-group of acompound of formula

wherein Y is as defined above, is first protected in a manner known perse, before reducing the carboxyl group to yield the correspondingalcohol of formula

wherein PC is a protective group and Y is as defined above. The alcoholof formula (6) is then converted to the corresponding compound offormula (7)

wherein LG is a leaving group, for example, chlorine, bromine, mesylateor tosylate, which is in turn converted to the corresponding sulfonicacid of formula (8)

using sodium sulphite or sodium thioacetate followed by hydrogenperoxide oxidation.

The compound of formula (8), after having been converted to thecorresponding methane sulfonic halide, for example by reaction withthionyl chloride, is reacted with an amine of formula

H₂N—R  (9),

wherein R is as defined above, to yield a compound of formula (4) inprotected form, which is finally deprotected. The above outlined stepsfrom the compound of formula (5) to the compound of formula (4) are allwell-known reactions which may be performed as disclosed in textbooks oforganic chemistry. The working examples further illustrate thereactions.

In the alternative, the compounds of formula (1) may be synthesized inanalogy to WO2010/020905, Scheme II on page 14.

In addition, a compound of formula (1), wherein A is a radical offormula

wherein R⁰ is as defined above, may be obtained by preparing first ofall a compound of formula

wherein PC is a protective group, by a process as described above,reducing the nitro group of the compound of formula (1a) in a mannerknown per se, for example catalytically with H₂/Rancy Nickel, andreacting the resulting diamine with an acid halide R⁰—C(O)Hal or with atrialkyl ortho-ester (AlkO)₃—C—R⁰, wherein Hal is Br or Cl, Alk is, forexample, ethyl and R⁰ is as disclosed above, in order to yield acompound of formula

after deprotection of the amine, wherein R and R⁰ are as disclosedabove.

The compounds of the present invention are Janus Kinase inhibitors(JAK-i) with efficacy, for example, against Janus Kinase-1 (JAK-1),Janus Kinase-2 (JAK-2), Janus Kinase-3 (JAK-3) and Tyrosine Kinase-2kinase (TYK-2), in particular JAK-1 or JAK-3. Accordingly, they areuseful as therapeutic agents for organ transplants, lupus, multiplesclerosis, rheumatoid arthritis, psoriasis, Type 1 diabetes andcomplications from diabetes, cancer, asthma, atopic dermatitis,autoimmune thyroid disorders, ulcerative colitis, inflammatory boweldiseases, Crohn's disease, Alzheimer's disease, leukemia,osteoarthritis, control of pruritus, chronic respiratory disease,keratoconjunctivitis and other indications whereimmunosuppression/immunomodulation would be desirable.

In particular it has turned out that the compounds of the presentinvention are safe and efficacious agents to control skin diseases,conditions or disorders including atopic dermatitis, eczema, psoriasis,scleroderma, pruritus and other pruritic conditions; allergic reactionsincluding allergic dermatitis in mammal including horse allergicdiseases such as bite hypersensitivity, summer eczema and sweet itch inhorses.

As the compounds of the present invention are JAK inhibitors withefficacy against JAK-I and JAK-3, they provide resolution of chronicpruritus and inflammation that would either persist in atopic dermatitisor slowly regress following removal of allergen or causative agent, suchas fleas in flea-allergic dermatitis.

Compounds of the present invention may be administered in apharmaceutically acceptable form either alone or in combination with oneor more additional agents which modulate a mammalian immune system orwith antiinflammatory agents. Examples are cyclosporin A aspirin,acetaminophen, ibuprofen, naproxen, piroxicam, and antiinflammatorysteroids (e.g. prednisolone or dexamethasone). These agents may beadministered as part of the same or separate dosage forms, via the sameor different routes of administration, and on the same or differentadministration schedules according to standard pharmaceutical practiceknown to one skilled in the art.

In one embodiment, the invention provides methods of treating orpreventing a disease, condition or disorder associated with JAK in asubject, such as a human or non-human mammal, comprising administeringan effective amount of one or more compounds described herein to thesubject. The JAK associated disease, condition or disorder can berelated to JAK-I, JAK-2, JAK-3, and/or TYK-2. Suitable subjects that canbe treated include domestic or wild animals, companion animals, such asdogs, cats, horses and the like; livestock including, cows and otherruminants, pigs, poultry, rabbits and the like; primates, for examplemonkeys, and rodents such as rats mice, gerbils guinea pigs and thelike. In one embodiment, the compound is administered in apharmaceutically acceptable form, optionally in a pharmaceuticallyacceptable carrier.

Another embodiment provides a method of inhibiting a JAK enzyme,including JAK-I, JAK-2, JAK-3 and/or Tyk-2, that includes contacting theJAK enzyme with either a non-therapeutic amount or a therapeuticallyeffective amount of one or more of the present compounds. Such methodscan occur in vivo or in vitro. In vitro contact can involve a screeningassay to determine the efficacy of the one or more compounds against aselected enzyme at various amounts or concentrations. In vivo contactwith a therapeutically effective amount of the one or more compounds caninvolve treatment of a described disease, disorder or condition orprophylaxis of organ transplant rejection in the animal in which thecontact occurs. The effect of the one or more compounds on the JAKenzyme and/or host animal can also be determined or measured. Methodsfor determining JAK activity are shown in the Examples part below.

In therapeutic use for treating disorders in a mammal (i.e. human andanimals), a compound of the present invention or its pharmaceuticalcompositions can be administered orally, parenterally, topically,rectally, transmucosally, or intestinally. Parenteral administrationsinclude indirect injections to generate a systemic effect or directinjections to the afflicted area. Topical administrations include thetreatment of skin or organs readily accessible by local application, forexample, eyes or cars. It also includes transdermal delivery to generatea systemic effect. The rectal administration includes the form ofsuppositories. The preferred routes of administration are oral andparenteral.

Pharmaceutical compositions of the present invention may be manufacturedby processes well known in the art, e.g., by means of conventionalmixing, dissolving, granulation, dragee-making, levigating, emulsifying,encapsulating, entrapping, lyophilizing processes or spray drying.Pharmaceutical compositions for use in accordance with the presentinvention may be formulated in conventional manner using one or morepharmaceutically acceptable carriers comprising excipients andauxiliaries, which facilitate processing of the active compound intopreparations, which can be used pharmaceutically. Proper formulation isdependent upon the route of administration chosen. Pharmaceuticallyacceptable excipients and carriers are generally known to those skilledin the art and are thus included in the instant invention.

The formulations of the invention can be designed to be short-acting,fast-releasing, long-acting, and sustained-releasing. Thus, thepharmaceutical formulations can also be formulated for controlledrelease or for slow release.

Pharmaceutical compositions suitable for use in the present inventioninclude compositions wherein the active ingredients are contained in anamount sufficient to achieve the intended purpose, i.e., control or thetreatment of disorders or diseases. More specifically, a therapeuticallyeffective amount means an amount of compound effective to prevent,alleviate or ameliorate symptoms/signs of disease or prolong thesurvival of the subject being treated.

The quantity of active component, which is the compound of thisinvention, in the pharmaceutical composition and unit dosage formthereof, may be varied or adjusted widely depending upon the manner ofadministration, the potency of the particular compound and the desiredconcentration. Determination of a therapeutically effective amount iswell within the capability of those skilled in the art. Generally, thequantity of active component will range between 0.01% to 99% by weightof the composition.

Generally, a therapeutically effective amount of dosage of activecomponent will be in the range of about 0.01 to about 100 mg/kg of bodyweight/day, preferably about 0.1 to about 10 mg/kg of body weight/day,more preferably about 0.3 to 3 mg/kg of body weight/day, even morepreferably about 0.3 to 1.5 mg/kg of body weight/day. It is to beunderstood that the dosages may vary depending upon the requirements ofeach subject and the severity of the disorders or diseases beingtreated. The desired dose may conveniently be presented in a single doseor as divided doses administered at appropriate intervals, for example,as two, three, four or more sub-doses per day. The sub-dose itself maybe further divided, e.g., into a number of discrete loosely spacedadministrations; such as multiple inhalations from an insufflator or byapplication of a plurality of drops into the eye. Also, it is to beunderstood that the initial dosage administered may be increased beyondthe above upper level in order to rapidly achieve the desired plasmaconcentration. On the other hand, the initial dosage may be smaller thanthe optimum and the daily dosage may be progressively increased duringthe course of treatment depending on the particular situation. Ifdesired, the daily dose may also be divided into multiple doses foradministration, e.g., two to four times per day.

The Examples further illustrate the invention.

EXAMPLE 1

This example illustrates the preparation of1-[trans-4-(imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)cyclohexyl]-N-methylmethanesulfonamide(Compound 22 in Table 1)

Step A

methyl trans-4-aminocyclohexanecarboxylate (0.19 g), acetonitrile (3 ml)and K₂CO₃ (0.407 g) were placed in a round bottomed flask with magneticagitator and heating bath. Benzyl bromide (0.29 ml) was added andreaction mixture was vigorously agitated at 25 to 30° C. for 3 hours.TLC (DCM/MeOH 8:1) revealed foil conversion of starting material andmonobenzylated amine presence. Inorganic precipitate was filtered off.Filtrate was evaporated. The residue was purified by chromatography onsilica gel (DCM/Hexane 1:1 to DCM/MeOH 10:1) to yield methyltrans-4-(dibenzylamino)cyclohexanecarboxylate as white needles (0.33 g).

Step B

methyl trans-4-(dibenzylamino)cyclohexanecarboxylate (0.22 g) wasdissolved in anhydrous THF (2.2 ml) and cooled to 0° C. Lithium aluminumhydride (0.125 g) was added portionwise over ca. 15 minutes. Whenfoaming ceased batch temperature was slowly increased and reactionaccelerated (exothermic). After 30 minutes sampled for TLC (DCM) and nostarting material was detected. Reaction mixture was quenched with waterand 10% NaOH. Phases were separated. Aqueous was extracted with methylr-butyl ether. The combined organic phases were washed with brine, driedover anhydrous sodium sulfate and concentrated in vacuo to yield[trans-4-(dibenzylamino)cyclohexyl]methanol as a colorless oilsolidifying upon standing (0.20 g). The crude product obtained was usedwithout further purification.

Step C

[trans-4-(dibenzylamino)cyclohexyl]methanol (7.74 g) andtriphenylphosphine (9.84 g) were dissolved in anhydrous THF (60 ml).Tetrabromomethane (12.44 g) was dissolved in THF (17.5 ml) and addeddropwise to the reaction mixture. The flask was cooled with water (˜10°C.) due to exothermicity. Precipitate appeared. After 1 hour sampled forTLC (DCM 100%) that revealed completed reaction. The solvent wasevaporated and the residue purified by chromatography on silica gel (DCM100%). Fractions containing product were combined and evaporated. Solidresidue was taken with hexane (30 ml) and cooled to 2 to 4° C. Theprecipitate was filtered off, rinsed with cold hexane and dried undervacuum to yield N,N-dibenzyl-trans-4-(bromomethyl)cyclohexanamine as awhite solid (9.2 g).

Step D

N,N-dibenzyl-trans-4-(bromomethyl)cyclohexanamine (5.0 g) was suspendedin isopropyl alcohol (10 ml). Na₂SO₃ (2.2 g) and KI (cat) were dissolvedin water (20 ml) and added to the suspension ofN,N-dibenzyl-4-(bromomethyl)cyclohexanamine in a pressure reactor. Itwas sealed and heated to 130° C. with good agitation. Reaction progresswas controlled by TLC (DCM 100%). When reaction was found to becompleted, solvents were evaporated and dried by azeotropic distillationwith toluene to yield [trans-4-(dibenzylamino)cyclohexyl]methanesulfonic acid. The crude product obtained was used withoutfurther purification.

Step E

Crude [trans-4-(dibenzylamino)cyclohexyl]methanesulfonic acid (4.5 g)was suspended in chloroform (75 ml) and cooled in an ice bath. Thionylchloride (19.4 ml) was added dropwise. Reaction mixture was agitated for20 minutes at RT then heated to 65° C. and agitated at 65° C. overnight.Solvent and excess of thionyl chloride were evaporated to afford[trans-4-(dibenzylamino)cyclohexyl]methanesulfonyl chloride. The crudeproduct obtained was used without further purification.

Step F

Crude [trans-4-(dibenzylamino)cyclohexyl]methanesulfonyl chloride (˜11.5mmol) was suspended in anhydrous THF (45 ml) and cooled to 0° C.Triethylamine (2.4 ml) was added followed by methylamine (2M in THF,11.5 ml). Reaction mixture was agitated at 0° C. for 1 hour, warmed toRT and held for 1 hour at RT. The solvent was evaporated and the residuewas taken with EtOAc (100 ml) and washed with NaHCO₃ (sat.). Aqueousphase was backwashed with EtOAc (50 ml). The combined organic phaseswere washed with water and brine (50 ml each), dried over magnesiumsulfate and concentrated in vacuo. The crude material was purified bychromatography on silica gel (DCM/MeOH 8:1). Fractions containingproduct were combined, concentrated in vacuo and triturated withDCM/hexane (1:1). Precipitate was filtered out, washed with coldDCM/hexane (1:1) and dried under vacuum to yield1-[trans-4-(dibenzylamino)cyclohexyl]-N-methyl-methanesulfonamide as awhite solid (1.67 g).

Step G

1-[trans-4-(dibenzylamino)cyclohexyl]-N-methyl-methanesulfonamide (1.6g) was dissolved (partially) m MeOH (16 ml). Ammonium formate (1.04 g)and wet 10% Pd/C (0.3 g) were added. Reaction mixture was heated toreflux with good agitation. Very stow conversion was encountered(TLC-DCM/MeOH 8:1). Solvent was topped up with THF (10 ml). Anotherportion of ammonium formate (1.04 g) was added followed by 10% Pd(OH)₂/C(0.3 g). Reaction mixture was resampled for TLC (DCM/McOH 8:1) after 1hour—full conversion was noted. The palladium catalyst was filtered outthrough a plug of Celite. The filter cake was washed with MeOH (2×20ml). Filtrates were combined and evaporated. Solid residue was takenwith methanol and evaporated (to remove remaining aromatic volatiles andammonium formate). This step was repeated twice to give1-(trans-4-aminocyclohexyl)-N-methyl-methanesulfonamide as an off-whitesolid (0.85 g).

Step H

To a suspension of 4-Chloro-7-azaindole (1.37 g), triethylamine (1.9 ml)and DMAP (0.11 g) in DCM (70 ml) wax added at RT benzenesulfonylchloride (1.3 ml). The reaction mixture was stirred at RT overnight. Thereaction mixture was diluted with DCM and was quenched with an aqueoussolution of HCl (1M, 70 ml). The organic phase was separated andextracted with a saturated solution of NaHCO₃ (70 ml), with water andwith a saturated aqueous solution of NaCl, dried over Na₂SO₄ andconcentrated in vacuo to yield1-(benzenesulfonyl)-4-chloro-pyrrolo[2,3-b]pyridine as a brown solid(2.65 g). The crude product obtained was used without furtherpurification.

Step I

Tetrabutylammonium nitrate (381 mg) dissolved in DCM (5 ml) was addeddropwise to a solution of1-(benzenesulfonyl)-4-chloro-pyrrolo[2,3-b]pyridine (292 mg) in DCM (5ml) under nitrogen at −10° C. Trifluoroacetic anhydride (180 μl) wasadded dropwise, stirred for 30 minutes at the same temperature and thenfor 4 hours at RT. Additional tetrabutylammonium nitrate (80 mg) andtrifluoroacetic anhydride (40 μl) were added and the reaction mixturewas stirred at room temperature overnight. Additional tetrabutylammoniumnitrate (380 mg) and trifluoroacetic anhydride (180 μl) were added andthe reaction mixture was stirred at room temperature for 3 hours. Afterdiluting with DCM, the reaction mixture was quenched with water. Theorganic phase was separated and extracted 3 times with water and oncewith a saturated aqueous solution of NaCl, dried over Na₂SO₄ andconcentrated in vacuo. Ille crude product was purified by chromatographyon silica gel (EtOAc/heptane 1:5) to yield1-(benzenesulfonyl)-4-chloro-5-nitro-pyrrolo[2,3-b]pyridine as a beigesolid (111 mg).

Step J

1-(benzenesulfonyl)-4-chloro-5-nitro-pyrrolo[2,3-b]pyridine (337 mg),1-(trans-4-aminocyclohexyl)-7V-methyl-methanesulfonamide (example 1,step G, 206 mg) and potassium carbonate (304 mg) were suspended indioxan/water 9:1 (10 ml). The resulting suspension was heated to 120° C.in a microwave oven. After 2 hours, the reaction mixture was sampled forHPLC/MS—no starting material visible; product was formed. The reactionmixture was concentrated in vacuo. THF (15 ml), EtOAc (45 ml) and water(30 ml) were added to the residue. The reaction mixture was stirred atRT for 30 minutes. The aqueous phase was separated and extracted twicewith a mixture of THF/EtOAc 1:3. The combined organic phases wereextracted with a saturated aqueous solution of NaCl, dried over Na₂SO₄and concentrated in vacuo. The crude product was purified bychromatography on silica gel (EtOAc/heptane 1:1 to EtOAc 100%) to yield1-[trans-4[[1-(benzenesulfonyl)-5-nitro-1H-pyrrolo[2,3-b]pyridin-4-yl]amino]cyclohexyl]-N-methyl-methanesulfonamideas a yellow foam (276 mg).

Step K

1-[trans-4-[[1-(benzenesulfonyl)-5-nitro-1H-pyrrolo[2,3-b]pyridin-4-yl]amino]cyclohexyl]-N-methyl-methanesulfonamide(254 mg) was dissolved in THF (20 ml). This solution was hydrogenatedfor 6 hours over a Raney-Nickel catalyst using the H-Cube® flow reactor(Full H₂-Mode, temperature=RT, flow rate=1 ml/min). The THF was removedunder vacuum. The residue was dissolved in EtOAc, dried over magnesiumsulfate and concentrated in vacuo to yield1-[trans-4-[[5-amino-1-(benzenesulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl]amino]cyclohexyl]-W-methyl-methanesulfonamideas a yellow resin (222 mg). The crude product obtained was used withoutfurther purification.

Step L

A mixture of1-[trans-4-[[5-amino-1-(benzenesulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl]amino]cyclohexyl]-N-methyl-methanesulfonamide(95 mg), triethyl orthoformate (80 μl) and p-toluenesulfonic acidmonohydrate (4 mg) in toluene (6 ml) was refluxed overnight.

After diluting with EtOAc, the reaction mixture was quenched with asaturated aqueous solution of NaHCO₃. The organic phase was separatedand was extracted once again with with a saturated aqueous solution ofNaHCO₃ and with a saturated aqueous solution of NaCl, dried over MgSO₄and concentrated in vacuo. The crude product was purified on asemi-preparative HPLC to yield1-[trans-4-(6-(phenylsulfonyl)-imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)cyclohexyl]-N-methylmethanesulfonamideas a colorless resin (50 mg).

Step M

A mixture of1-[trans-4-(6-(phenylsulfonyl)-imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)cyclohexyl]-N-methylmethanesulfonamide(68 mg) and lithium hydroxide (14 mg) in isopropanol/water 1:1 (1 ml)was stirred at 40° C. overnight. After 20 hours, the reaction mixturewas heated to 50° C. and stirred at 50° C. for one day.

Additional lithium hydroxide (14 mg) was added. The reaction mixture washeated to 60° C. and stirred at 60° C. for one day. An aqueous solutionof HCl 37% (120 μl) was added until pH=5 was reached and then asaturated aqueous solution of NaHCO₃ (100 μl) was added until pH=8 wasreached. Isopropanol was evaporated. EtOAc/THF (2-3 ml) was added to theresidue and the resulting suspension was stirred at room temperature. Anaqueous solution of potassium carbonate (2 M, 2-3 ml) was added untilpH=12 was reached. The suspension was further stirred at roomtemperature and the precipitate was filtered off rinsed with water anddried under vacuum to yield1-[trans-4-(imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)cyclohexyl]-N-methylmethanesulfonamide(13 mg, compound 22 in Table 1). MS (HPLC/MS): 348 (MH⁺). Retentiontime: 1.70 min.

EXAMPLE 2

This example illustrates the preparation ofN-methyl-1-[trans-4-[(5-nitro-1H-pyrrolo[2,3-b]pyridin-4-yl)oxy]cyclohexyl]methanesulfonamide(Compound 25 in Table 1)

Step A

Ethyl trans-4-hydroxycyclohexanecarboxylate (10 g), diisopropylamine (21ml), benzyl bromide (10 ml) and sodium iodide (0.9 g) were mixedtogether and heated in a sealed tube at 120° C. overnight. Afterdiluting with EtOAc, the reaction mixture was quenched with water. Theaqueous phase was separated and extracted twice with EtOAc. The combinedorganic phases were dried over anhydrous MgSO₄ and concentrated invacuo. The crude product was purified by chromatography on silica gel(hexane/EtOAc 9:1) to yield ethyltrans-4-(benzyloxy)cyclohexanecarboxylate as a yellow oil (16.0 g).

Step B

Ethyl trans-4-(benzyloxy)cyclohexanecarboxylate (16.0 g) was suspendedin dry THF while cooling in an ice bath. Lithium aluminum hydride (5.2g) was added portionwise. After addition reaction mixture was heated at60° C. for 4 hours. After that time the reaction mixture was cooled to0° C. and EtOAc (30 ml) followed by water (30 ml) were added. Theresulting inorganic salts were filtered off through a pad of Celite.Phases were separated. Aqueous was extracted with EtOAc. The combinedorganic phases were dried over anhydrous MgSO₄ and concentrated in vacuoto yield (trans-4-benzyloxycyclohexyl)methanol as a light yellow solid(14.2 g). The crude product obtained was used without furtherpurification.

Step C

(trans-4-benzyloxycyclohexyl)methanol (14.2 g) was suspended in dry THFwhile cooling in an ice bath. Triphenylphosphine (22.32 g) was added.The resulting solution was stirred at 0° C. for 10 minutes, thentetrabromomethane (28.22 g) was added portionwise and the slurry wasallowed to reach room temperature. After 24 hours of stirring the whiteprecipitate was filtered and washed with THF followed by EtOAc. Thefiltrate was evaporated under reduced pressure and purified by columnchromatography on silica gel (hexane/EtOAc 9:1) to yield benzyltrans-4-(bromomethyl)cyclohexyl ether as a yellow solid (17.0 g).

Step D

benzyl trans-4-(bromomethyl)cyclohexyl ether (8.0 g) was dissolved inisopropanol (100 ml) and sodium sulphite (7.12 g) in water (100 ml) wasadded. The reaction mixture was then stirred vigorously while heated at100° C. overnight. After cooling to room temperature, the reactionmixture was concentrated to give a white solid. Methanol was added andthe mixture was stirred at RT for 3 hours, then the precipitate wasfiltered off, rinsed with methanol, the filtrate was evaporated to yield(trans-4-benzyloxycyclohexyl)methanesulfonic acid as a white solid (9.5g).

Step E

(trans-4-benzyloxycyclohexyl)methanesulfonic acid (1.0 g) was suspendedin a freshly distilled chloroform (50 ml) while cooling m an ice bath.Dry DMF (3-5 drops) was added. The resulting solution was stirred at 0°C. for 10 minutes, then thionyl chloride (0.52 ml) was added drop wise.The mixture was stirred at this temperature for 15 minutes, 30 minutesat room temperature and overnight at 45° C. After cooling down, thesolvent was evaporated; dry DCM was added and evaporated to remove theresidual thionyl chloride. This procedure was repeated twice to yield(trans-4-benzyloxycyclohexyl)methanesulfonyl chloride as a yellow oil(1.0 g).

Step F

(trans-4-benzyloxycyclohexyl)methanesulfonyl chloride (1.0 g) wassuspended m dry DCM (50 ml) while cooling in an ice bath, methylamine(2M in THF, 5.0 ml) was added drop wise. Subsequently, the reactionmixture was allowed to reach room temperature and stirred at thistemperature overnight. After that time, the solvent was evaporated andpurified by column chromatography on silica gel (hexane/EtOAc 9:1) toyield 1-(trans-4-benzyloxycyclohexyl)-N-methyl-methanesulfonamide as alight yellow solid (0.56 g).

Step G

1-(trans-4-benzyloxycyclohexyl)-N-methyl-methanesulfonamide (3.7 g) wassuspended in methanol, Pd(OH)₂ (1.75 g) was added. Subsequently, thereaction was continued in a Parr apparatus for 12 hours. Then, thecatalyst was filtered off through a pad of Celite. Filtrate wasevaporated, washed with Et₂O and dried under vacuum to yield1-(trans-4-hydroxycyclohexyl)-N-methyl-methanesulfonamide as a whitesolid (2.44 g).

Step H

Sodium hydride (60% in mineral oil, 0.09 g) was added under nitrogen toa solution of 1-(trans-4-hydroxycyclohexyl)-N-methyl-methanesulfonamide(0.40 g) in DMF (19 ml). After 30 minutes at room temperature,2-(trimethylsilyl)ethoxymethyl chloride was added over 15 minutes to thereaction mixture. After 18 hours at room temperature, the reactionmixture was diluted with EtOAc and quenched with an aqueous solution ofsodium phosphate (1 M). The aqueous phase was separated and extractedtwice with EtOAc. The combined organic phases were washed with water andtwice with brine, dried over anhydrous magnesium sulfate andconcentrated in vacuo to yield1-(trans-4-hydroxycyclohexyl)-N-methyl-N-(2-trimethylsilylethoxymethyl)methanesulfonamideas a yellow oil (0.66 g). The crude product obtained was used withoutfurther purification.

Step I

Sodium hydride (60% in mineral oil, 0.07 g) was added under nitrogen toa solution of1-(trans-hydroxycyclohexyl)-N-methyl-N-(2-trimethylsilylethoxymethyl)methanesulfonamide(0.49 g) in DMF (10 ml). After 30 minutes at room temperature,1-(benzenesulfonyl)-4-chloro-5-nitro-pyrrolo[2,3-b]pyridine (example 1,step I, 0.49 g) in DMF (5 ml) was added over 15 minutes to the reactionmixture. After three days at room temperature, EtOAc was added mid thereaction mixture was poured onto water. The aqueous phase was separatedand extracted with EtOAc. The combined organic phases were washed twicewith water, twice with an aqueous sodium hydroxide solution (2 N) andtwice with brine, dried over anhydrous magnesium sulfate andconcentrated in vacuo. The crude product was purified on asemi-preparative HPLC to yieldN-methyl-1-[trans-4-[(5-nitro-1H-pyrrolo[2,3-b]pyridin-4-yl)oxy]cyclohexyl]-N-(2-trimethylsilylethoxymethyl)methanesulfonamideas a yellow solid (0.12 g).

Step J

A solution ofN-methyl-1-[trans-4-[(5-nitro-1H-pyrrolo[2,3-b]pyridin-4-yl)oxy]cyclohexyl]-N-(2-trimethylsilylethoxymethyl)methanesulfonamide(120 mg) in AcOH (10 ml) and water (5 ml) was stirred at 70° C. for 1hour under nitrogen. The reaction mixture was cooled to room temperatureand poured onto water. An aqueous solution of NaOH (4N) was added untilpH=7-8 was reached. The aqueous phase was extracted twice with EtOAc.The Combined organic phases were washed with water, twice with anaqueous sodium hydroxide solution (2 N) and with brine, dried overanhydrous magnesium sulfate and concentrated in vacuo. The residue wasrecrystallized from DCM/EtOAc (9:1) to yieldN-methyl-1-[trans-4-[(5-nitro-1H-pyrrolo[2,3-b]pyridin-4-yl)oxy]cyclohexyl]methanesulfonamide(50 mg, compound 25 in Table 1). MS (HPLC/MS): 369 (MH⁺). Retentiontime: 1.12 min.

EXAMPLE 3

This example illustrates the preparation of1-[trans-4-[(5-cyano-1H-pyrrolo[2,3-b]pyridin-4-yl)-methyl-amino]cyclohexyl]-N-methyl-methanesulfonamide(Compound 26 in Table 1).

Step A

Sodium bis(2-methoxyethoxy)aluminum hydride solution (Red-Al®, 65% intoluene, 183 ml) is added over 60 minutes to a solution oftrans-4-(tert-butoxycarbonylamino)cyclohexanecarboxylic acid (24.3 g) intoluene (250 ml) at 0° C. The reaction mixture was then heated to 130°C. and stirred at this temperature for 1 hour. After cooling to 0° C., asaturated aqueous solution of sodium sulfate (195 ml) was added dropwise. The reaction mixture was then filtered off through a Hyflo fitter.The fitter cake was rinsed with DCM (150 ml) and water (24 ml). Theaqueous phase was separated and extracted twice with DCM (2×150 ml). Thecombined organic phases were dried over anhydrous magnesium sulfate andconcentrated in vacuo to yield [trans-4-(methylamino)cyclohexyl]methanolas white crystals (12.5 g). The crude product obtained was used withoutfurther purification.

Step B

Benzoyl chloride (8.8 ml) was added dropwise to an emulsion of sodiumhydrogencarbonate (12.6 g) in water (50 ml) and[trans-4-(methylamino)cyclohexyl]methanol (10.9 g) in DCM (50 ml) at 0°C. Subsequently, the reaction mixture was allowed to reach roomtemperature and stirred at this temperature for 3 hours. The reactionmixture was diluted with water (150 ml) and with DCM (200 ml). Theorganic phase was separated, dried over anhydrous magnesium sulfide andconcentrated in vacuo to yieldN-[trans-4-(hydroxymethyl)cyclohexyl]-N-methylbenzamide as beigecrystals (17.2 g). The crude product obtained was used without furtherpurification.

Step C

Triethylamine (11 ml), DMAP (0.43 g) and p-toluenesulfonyl chloride(13.2 g) were added to a solution ofN-[trans-4-(hydroxymethyl)cyclohexyl]-N-methylbenzamide (17.0 g) in DCM(250 ml). After 3 hours at room temperature, the reaction mixture wasquenched with water (150 ml). The organic phase was separated, driedover anhydrous magnesium sulfate and concentrated in vacuo. The crudeproduct was purified by chromatography on silica gel (EtOAc/heptane 1:1)to yield [trans-4-[benzoyl(methyl)amino]cyclohexyl]methyl4-methylbenzenesulfonate as white crystals (19.5 g).

Step D

Potassium thioacetate (6.3 g) was added to a suspension of[trans-4-[benzoyl(methyl)amino]cyclohexyl]methyl4-methylbenzenesulfonate (19.5 g) in DMSO (66 ml) at RT. The reactionmixture was heated to 55° C. and stirred at 55° C. for 3 hours. Aftercooling down at room temperature, the reaction mixture was diluted withEtOAc (200 ml) and quenched with an aqueous solution of NaHCO₃ (0.1 M,300 ml). The aqueous phase was separated and extracted twice with EtOAc(2×200 ml). The combined organic phases were washed with water andbrine, dried over anhydrous magnesium sulfate and concentrated in vacuo.The residue (17 g, light yellow crystals) was dissolved in formic acid(73 ml) and the reaction mixture was heated to 25-35° C. A hydrogenperoxide solution (30% in water, 25 ml) was added over 60 minutes atthis temperature. Subsequently, the reaction mixture was cooled down atroom temperature and stirred at RT for 15 minutes. After cooling down at0° C., the reaction mixture was quenched with an aqueous solution ofsodium metabisulfite (33%, 27 ml). An aqueous solution of NaOH (33%, 121ml) was then added at 0° C. until pH=5 was reached and the reactionmixture was stirred at RT overnight. The reaction mixture wasconcentrated in vacuo. The residue obtained was taken up in water (160ml) and 2-propanol (40 ml) and stirred at 45° C. Subsequently,2-propanol was evaporated and an aqueous solution of HCl (2 M, 10 ml)was added. The suspension was stirred at 0° C. and filtered off. Theprecipitate was dried under vacuum over Sicapent® to yield[trans-4-[benzoyl(methyl)amino]cyclohexyl]methanesulfonic acid as awhite solid (31.0 g) used in the next step without further purification.

Step E

[trans-4-[benzoyl(methyl)amino]cyclohexyl]methanesulfonic acid (10.1 g)was suspended in DCM (40 ml) with addition of a few drops of DMF. Thereaction mixture was cooled down at 0° C. and thionyl chloride (4.7 ml)was added drop wise over 10 minutes. The reaction mixture was thenrefluxed for 6 hours. After cooling down at 0° C., additional thionylchloride (4.7 ml) was added drop wise over 10 minutes. The reactionmixture was then refluxed overnight. After that time, the mixture wascooled to room temperature and concentrated nearly to dryness in vacuo.Dry toluene was added to the residue and removed under reduced pressureto ensure the removal of any unreacted thionyl chloride. The residue wastaken up in THF (40 ml) and methylamine was added (2 M in THF, 47 ml).The reaction mixture was stirred at RT overnight. The resultingsuspension was filtered and rinsed with THF. The filtrate was evaporatedto afford a light brown resin (2.6 g). The precipitate was suspended inDCM (20 ml) with addition of a few drops of DMF. Thionyl chloride (2.4ml) was added dropwise over 10 minutes. The reaction mixture was thenrefluxed overnight. After 16 hours under reflux, additional thionylchloride (2.4 ml) was added dropwise over 10 minutes. The reactionmixture was then refluxed overnight. After that time, the mixture wascooled to room temperature and concentrated nearly to dryness in vacuo.Dry toluene was added to the residue and removed under reduced pressureto ensure the removal of any unreacted thionyl chloride. The residue wastaken up in THF (20 ml) and methylamine was added (2 M in THF, 24 ml).The reaction mixture was stirred at RT for 4 hours, then was filteredand rinsed with THF. The filtrate was evaporated to afford a light brownresin (2.0 g). The combined light brown resins (4.6 g) were purified bychromatography on silica gel (McOH/DCM 1:49 to 1:19) to yieldN-methyl-N-[trans-4-(methylsulfamoylmethyl)cyclohexyl]benzamide as beigecrystals (2.18 g).

Step F

N-methyl-N-[trans-4-(methylsulfamoylmethyl)cyclohexyl]benzamide (875 mg)was taken up in an aqueous solution of HCl (6 M, 10 ml) and the reactionmixture was refluxed for two days. After cooling down at roomtemperature, the resulting suspension was filtered. The filtrate wasconcentrated in vacuo to yieldN-methyl-1-[trans-4-(methylamino)cyclohexyl]methanesulfonamidehydrochloride as beige crystals (710 mg). The crude product obtained wasused without further purification.

Step G

N-methyl-1-[trans-4-(methylamino)cyclohexyl]methanesulfonamidehydrochloride (129 mg), 4-Chloro-1H-pyrrolo[2,3-b]pyridine-carbonitrile(89 mg) and potassium carbonate (225 mg) were suspended in water/dioxane1:9 (4.5 ml). The resulting suspension was heated to 160° C. in amicrowave oven for 8 hours. The reaction mixture was concentrated invacuo. The residue was taken up in THF (10 ml) and a mixture ofwater/aqueous saturated solution of NaCl 1:1 (20 ml) was added. Themixture was stirred at RT. The aqueous phase was separated and extractedtwice with THF (2×10 ml). The combined organic phases were washed withbrine, dried over anhydrous magnesium sulfate and concentrated in vacuo.The residue was suspended in acetonitrile, stirred at RT, filtered andrinsed with acetonitrile. The filtrate was concentrated in vacuo and waspurified on a semi-preparative HPLC to yield1-[trans-4-[(5-cyano-1H-pyrrolo[2,3-b]pyridin-4-yl)-methyl-amino]cyclohexyl]-N-methyl-methanesulfonamide(22 mg, compound 26 in Table 1). MS (HPLC/MS): 362 (MH⁺). Retentiontime: 2.19 min.

EXAMPLE 4

This example illustrates the preparation ofN-methyl-1-[trans-4-[methyl(9H-purin-6-yl)amino]cyclohexyl]methanesulfonamide(Compound 2 in Table 1).

Step A

N-methyl-1-[trans-4-(methylamino)cyclohexyl]methanesulfonamidehydrochloride (example 3, step F, 140 mg), 6-chloropurine (59 mg) andtriethylamine (0.26 ml) were dissolved in n-butanol (4 ml). The reactionmixture was heated to 140° C., stirred at 140° C. overnight and heatedto 150° C. in a microwave oven for 1 hour. The reaction mixture wasconcentrated in vacuo. The residue was taken up in THF (3 ml) and amixture of water/aqueous saturated solution of NaCl 1:1 (4 ml) wasadded. The mixture was stirred at RT. The aqueous phase was separatedand extracted twice with THF (2×3 ml). The combined organic phases werewashed with brine, dried over anhydrous magnesium sulfate andconcentrated in vacuo. The residue was purified first by chromatographyon Isolute®NH₂ (DCM 100%, McOH/DCM 1:49 to 1:19), then on asemi-preparative HPLC and finally by chromatography on Isolute®NH₂(MeOH/DCM 1:24) to yieldN-methyl-1-[trans-4-[methyl(9H-purin-6-yl)amino]cyclohexyl]methanesulfonamide(22 mg, compound 2 in Table 1). MS (HPLC/MS): 339 (MH⁺). Retention time:1.87 min.

EXAMPLE 5

This example illustrates the preparation of1-[4-[(2-amino-5-methyl-pyrimidin-4-yl)-methyl-amino]cyclohexyl]-N-methyl-methanesulfonamide(Compound 29 in Table 1).

Step A

N-methyl-1-[trans-4-(methylamino)cyclohexyl]methanesulfonamidehydrochloride (example 3, step F, 129 mg),4-chloro-5-methyl-pyrimidin-2-amine (72 mg), Cs₂CO₃ (390 mg), Pd(OAc)₂(11 mg) and RuPhos (47 mg) were dissolved in tert-butanol (1 ml) in aschlenk tube. The reaction mixture was heated to 85° C., stirred at 85°C. overnight. The reaction mixture was taken up in THF (10 ml) andfiltered over celite. The filtrate was concentrated in vacuo. Theresidue was purified on a semi-preparative HPLC to yield1-[4-[(2-amino-5-methyl-pyrimidin-4-yl)-methyl-amino]cyclohexyl]-N-methyl-methanesulfonamide(Compound 29 in Table 1). MS (HPLC/MS): 328 (MH⁺). Retention time: 1.82min.

EXAMPLE 6

This example illustrates the preparation of1-[trans-4-((2-methoxymethyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)cyclohexyl]-N-methylmethanesulfonamide(Compound 35 in Table 1).

Step A

A mixture of1-[trans-4-[[5-amino-1-(benzenesulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl]amino]cyclohexyl]-N-methyl-methanesulfonamide(Example 1, step K) (212 mg), methoxyacetyl chloride (40 μl) andtriethylamine (70 μl) in methylenchloride (5 ml) was stirred an hour atroom temperature. The reaction mixture is concentrated in vacuo. Thecrude is taken up in acetic acid (2 mL) and heated at 100° C. for 3 h ina microwave. After diluting with EtOAc, the reaction mixture wasquenched with a saturated aqueous solution of NaHCO₃. The organic phasewas separated and was extracted once again with with a saturated aqueoussolution of NaHCO₃ and with a saturated aqueous solution of NaCl, driedover MgSO₄ and concentrated in vacuo. The crude product was purified ona semi-preparative HPLC to yield1-[trans-4-<(2-methoxymethyl)-6-(phenylsulfonyl)-imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-ylCyclohexyl]-N-methylmethanesulfonamide as a colorless resin.

Step B

1-[trans-4-((2-methoxymethyl)-6-(phenylsulfonyl)-imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)cyclohexyl]-N-methylmethanesulfonamideis deprotected using a similar procedure as described in Example 1, step1, to yield1-[trans-4-((2-methoxymethyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)cyclohexyl]-N-methylmethanesulfonamide(26 mg, compound 35 in Table 1). MS (HPLC/MS): 392 (MH⁺). Retentiontime: 1.89 min.

Analysis of the purified samples is m each case done using a WatersAutopurification (HPLC/MS) system with a reversed phase column (DaisogelSP-120-ODS-AP 5 μm, 150×3 mm) from Bischoff, Leonberg, Germany. Thesamples are characterized by m/z and retention time. The above-givenretention times relate in each case to the use of a solvent systemcomprising two different solvents, solvent A: H₂O+0.01% HCOOH, andsolvent B: CH₃CN+0.01% HCOOH). Said two solvents A and B are employed ata flow rate of 2.00 ml/min with a time-dependent gradient as given inthe Table:

Method A: column Daisogel SP-120-ODS-AP 5 μm, 150×3 mm) from Bischoff,Leonberg, Germany, flow rate of 2.00 mL/min with a time-dependentgradient as given in Table 1:

TABLE 1 Time [min] A [%] B [%] 0.5 90 10 1.0 74 26 1.5 60 40 2.0 47 532.5 36 64 3.0 26 74 3.5 19 81 4.0 13 87 4.25 10 90 4.5 8 92 4.75 7 935.0 6 94 5.5 5 95 6.5 5 95Method B: column Waters XTerra MS C18 5 μm, 50×4.6 mm (Waters), flowrate of 3.00 mL/min with a time-dependent gradient as given in Table 2

TABLE 2 Time [min] A [%] B [%] 0 90 10 0.5 90 10 2.5 5 95 2.8 5 95 2.990 10 3.0 90 10The substances named in the following Table 3 are prepared analogouslyto the above-described methods. The following physical data are obtainedaccording to the above-described HPLC/MS characterization process.

TABLE 3 Ex. R_(t) [min] Physical No. Compound of formula m/z: [M + H⁺](Method) state 1

353 1.72 (A) solid 2

339 1.87 (A) solid 3

339 1.80 (A) solid 4

415 3.80 (A) solid 5

429 3.50 (A) solid 6

325 1.38 (B) solid 7

392 1.80 (A) resin 8

341 2.08 (A) solid 9

355 2.08 (A) solid 10

348 2.26 (A) resin 11

300 1.71 (A) solid 12

314 1.83 (A) solid 13

378 1.84 (A) solid 14

365 2.53 (A) solid 15

324 1.95 (A) solid 16

314 1.89 (A) resin 17

379 2.67 (A) solid 18

382 2.38 (A) solid 19

342 1.33 (B) solid 20

366 1.37 (B) oil 21

390 2.26 (A) oil 22

348 1.70 (A) solid 23

376 2.28 (A) solid 24

362 2.72 (A) solid 25

369 1.12 (B) solid 26

362 2.19 (A) solid 27

348 2.75 A) solid 28

328 1.95 (A) foam 29

328 1.82 (A) foam 30

368 2.50 (A) solid 31

338 0.29 (B) foam 32

363 1.67 (A) solid 33

362 1.71 (A) solid 34

378 1.63 (A) solid 35

392 1.89 (A) foam

Measurement of the Effect on the JAK Enzyme

All four kinases of the JAK/TYK-kinase family were used as purifiedrecombinant GST-fusion proteins, containing the active kinase domains.GST-JAK1(866-1154), GST-JAK3(811-1124), and GST-TYK2(888-1187) wereexpressed and purified by affinity chromatography at the EPK biologyunit.

The kinase assays were based on the Caliper mobility shift assay usingthe LabChip 3000 systems. This technology is similar to capillaryelectrophoresis and uses charge driven separation of substrate andproduct in a microfluidic chip.

All kinase reactions were performed in 384 well microtiter plates in atotal reaction volume of

18 μl. The assay plates were prepared with 0.1 μl per well of testcompound in the appropriate test concentration, as described under thesection “preparation of compound dilutions”. The reactions were startedby combining 9 μl of substrate mix (consisting of peptide and ATP) with9 μl of kinase dilution. The reactions were incubated for 60 minutes at30° C. and stopped by adding 70 μl of stop buffer (100 mM Hepes, 5%DMSO, 0.1% Coating reagent, 10 mM EDTA, 0.015% Brij 35).

Fluorescently labeled synthetic peptides were used as substrates in allreactions. A peptide derived from the sequence of IRS-1 (IRS-1 peptide,FITC-Ahx-KKSRGDYMTMQIG-NH2) was used for JAK1 and TYK2 and a peptidenamed JAK3tide (FITC-GGEEEEYFELVKKKK-NH2) for JAK3. Specific assayconditions are described in Table 4:

TABLE 4 Assay conditions of individual kinase assays Kinase JAK1 JAK3TYK2 Buffer 50 mM Hepes 50 mM Hepes 50 mM Hepes pH 7.5, pH 7.5, pH 7.5,0.02% Tween 0.02% Tween 0.02% Tween 20, 1 mM DTT, 20, 1 mM DTT, 20, 1 mM0.02% BSA, 0.02% BSA, DTT, 12 mM MgCI2 1.5 mM MgCI2 0.02% BSA, 9 mMMgCI2 DMSO 0.6% 0.6% 0.6% Kinase conc. 50 nM 6 nM 40 nM Substrate 5 μM 2μM 5 μM peptide conc. ATP conc. 40 μM 80 or 18 μM 30 μM

The terminated reactions were transferred to the Caliper LabChip 3000reader and the turnover of each reaction was measured by determining thesubstrate/product ratio.

Preparation of Compound Dilutions

Test compounds were dissolved in DMSO (10 mM) and transferred into 1.4mL flatbottom or V-shaped Matrix tubes carrying a unique 2D matrix chipby individual compound hubs. The numbers of these chips weredistinctively linked to the individual compound identification numbers.The stock solutions were stored at −20° C. if not used immediately. Forthe test procedure the vials were defrosted and identified by a scannerwhereby a working sheet was generated that guided the subsequent workingsteps. Compound dilutions were made in 96 well plates. This formatenabled the assay of maximally 40 individual test compounds at 8concentrations (single points) including 4 reference compounds. Thedilution protocol included the production of pre-dilution plates, masterplates and assay plates:

Pre-dilution plates: 96 polypropylene well plates were used aspre-dilution plates. A total of 4 pre-dilution plates were preparedincluding 10 test compounds each on the plate positions A1-A10, onestandard compound at A11 and one DMSO control at A12. All dilution stepswere done on a HamiltonSTAR robot.

Master plates: 1OO μI of individual compound dilutions includingstandard compound and controls of the 4 “pre-dilution plates” weretransferred into a 384 “master plate” including the followingconcentrations 1,820, 564, 182, 54.6, 18.2, 5.46, 1.82 and 0.546 μM,respectively in 90% of DMSO.

Assay plates: Identical assay plates were then prepared by pipetting 100nL of compound dilutions of the master plates into 384-well “assayplates”. In the following the compounds were mixed with 9 μl of assayscomponents plus 9 μl enzyme corresponding to a 1:181 dilution stepsenabling the final concentration of 10, 3.0, 1.0, 0.3, 0.1, 0.03, 0.01and 0.003 μM, respectively. The preparation of the master plates werehandled by the Matrix PlateMate Plus robot and replication of assayplates by the HummingBird robot.

On the basis of this study, a compound of the invention showstherapeutic efficacy especially against disorder dependant on proteinkinase, especially proliferative diseases mediated by JAK/TYK kinaseactivity.

Ex. JAK1/IC50 JAK3/IC50 TYK2/IC50 No. (μM) (μM) (μM) 1 0.838 7.928 5.5132 0.582 1.886 4.593 3 >10 >10 >10 4 >10 >10 >10 5 >10 >10 >106 >10 >10 >10 7 >10 >10 >10 8 >10 >10 >10 9 >10 >10 >10 10 >10 >10 >1011 >10 >10 >10 12 >10 >10 >10 13 2 >10 6 14 >10 >10 >10 15 >10 >10 >1016 >10 >10 >10 17 >10 >10 >10 18 0.021 1.2 0.39 19 >10 >10 >1020 >10 >10 >10 21 >10 >10 >10 22 0.01 0.26 0.13 23 >10 >10 >10 240.93 >10 8.5 25 0.071 3.6 1.1 26 0.034 3 0.84 27 1.8 >10 7.8 28 0.55 >103.2

1.-20. (canceled)
 21. A compound of formula

wherein R is C₁-C₄-alkyl or C₃-C₅-cycloalkyl; and A is a radical offormula j

wherein R⁰ is H, C₁-C₄-alkyl, hydroxy-C₁-C₄-alkyl,C₁-C₄-alkoxy-C₁-C₄-alkyl or NH₂; or a veterinary acceptable saltthereof.
 22. The compound of claim 1 which is1-[trans-4-((2-amino)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)cyclohexyl]-N-methylmethanesulfonamideand can be structurally represented as

or a veterinary acceptable salt thereof.
 23. A pharmaceuticalcomposition comprising a compound according to claim 21 or a veterinaryacceptable salt thereof, and a pharmaceutical acceptable carrier.
 24. Amethod of treating atopic dermatitis, eczema, psoriasis, scleroderma,pruritus and other pruritic conditions, or allergic dermatitis, themethod comprising administering to a mammal in need thereof an effectiveamount of a compound according to claim 21 or a veterinary acceptablesalt thereof.
 25. The method according to claim 24, wherein the mammalis a dog.